LbLb?ID EU855183; SV 1; circular; other DNA; STD; SYN; 10466 BP. AC EU855183; DT 03-AUG-2008 (Rel. 96, Created) DT 03-AUG-2008 (Rel. 96, Last updated, Version 1) DE Cloning vector pAW8_TAP, complete sequence. KW . OS Cloning vector pAW8_TAP OC other sequences; artificial sequences; vectors. RN [1] RP 1-10466 RX DOI; 10.1016/j.gene.2007.09.024. RX PUBMED; 18054176. RA Watson A.T., Garcia V., Bone N., Carr A.M., Armstrong J.; RT "Gene tagging and gene replacement using recombinase-mediated cassette RT exchange in Schizosaccharomyces pombe"; RL Gene 407(1-2):63-74(2008). RN [2] RP 1-10466 RA Watson A.T.; RT ; RL Submitted (27-JUN-2008) to the EMBL/GenBank/DDBJ databases. RL Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton, RL East Sussex BN1 9RQ, UK FH Key Location/Qualifiers FT source 1..10466 FT /organism="Cloning vector pAW8_TAP" FT /mol_type="other DNA" FT /db_xref="taxon:550443" FT misc_feature 1..3873 FT /note="Cre-expression plasmid pAW8 backbone" FT misc_recomb 3874..3907 FT /note="loxP" FT CDS 3916..4467 FT /codon_start=1 FT /transl_table=11 FT /product="twin affinity purification tag" FT /note="TAP tag" FT /protein_id="ACF75949.1" FT /translation="MKRRWKKNFIAVSAANRFKKISSSGALDYGTPTTASENLYFQGEL FT KTAALAQHAIKADAQQNNFNKDQQSAFYEILNMPNLNEAQRNGFIQSLKDDPSQSTNVL FT GEAKKLNESQAPKADNNFNKEQQNAFYEILNMPNLNEEQRNGFIQSLKDDPSQSANLLS FT EAKKLNESQAPKADNKFNKE" FT terminator 4476..4704 FT /note="Saccharomyces cerevisiae ADH1" FT gene 5094..5903 FT /gene="kanR" FT CDS 5094..5903 FT /codon_start=1 FT /transl_table=11 FT /gene="kanR" FT /product="kanamycin and geneticin G418 resistance protein" FT /protein_id="ACF75950.1" FT /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP FT DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FT FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FT FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD FT RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" FT misc_recomb 6151..6184 FT /note="loxM3" FT misc_feature 6185..10466 FT /note="Cre-expression plasmid pAW8 backbone" SQ Sequence 10466 BP; 3004 A; 2247 C; 2159 G; 3056 T; 0 other; caaaaagtta ggtgtaacag aaaatcatga aactatgatt tctaatttat atattggagg 60 attttctcta aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg accatttgat 120 ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa 180 ttttactctg tcagaaacgg ccttaacgac gtagtcgatc cggacgatgg ccaaccttcc 240 aattcattaa atcgagtcct ccatcagatg atttacaagg ttttgcatag aatccctaag 300 taagagaagt tgttagtgat ttcttcaagt ttactaaaat ttacatacaa atggatgatc 360 tcttctccat tgtttccttt cttcttgcaa acgcgtttta caaagagatg acatgctatt 420 ttctgattat ttttttctat agttttctat tttgtacgtc cttgggagtc cgaaatgtaa 480 aatcggggta tggttagtag cggcagcgtt cattatggta tataaaagat aaaaagtagg 540 ggcaagcgag acagaatggg atacaagggc atcgtctata attatagcta aaaattgtat 600 tttaatttgt attttttgta attttatttc tcattgtttt acttaaaatg aatgcgaatt 660 agagaaaact tcaacgaaat gtcaaaataa gctcagcaaa atatacaatt ttagggaaag 720 cgatcagcaa cttattctcc ggtgtagatt ctttttcttc ataaattcca acaatgtaac 780 tgccaattga tttttctcta gatgctaata aattgaaact aatgaatata ataacacagg 840 ttttgataaa ttgtaaactc ttactcaaaa aaagaatgtt aaaactaatt taaccttttt 900 ataaaggctc atggcgttaa agaggatagt aaaaattcta caatttttta atttaatgtc 960 acaattgtct tgcgaatata attctattaa ttttcgctaa ctgctgtttt agtactatta 1020 ctttcaatat gccttatatg caactttaat tcatgaagaa tgcaatattt ttgcaatctt 1080 tcttgtactg agcgcttttt atcatttagt actcattatt taatttttca gtaaagggca 1140 ataaataaat ttgtagaaga tgcaatgtaa tctcctctat ccttttgctc atatgtttat 1200 gagtatacct agtctagaaa ggcttgtatt taaaatatta ttcaataaaa attcacaatt 1260 tttacgacat gtgctaccat tcacttaact tcctgattat aaaattggtt cgtttatact 1320 aattacttaa gtacctttaa ctaaacaaaa tgcctatata tatattaatt tacaatgagt 1380 gtcagataag tcactatgtc cgagtggtta aggagttaga ctcgccatcg atggtatggt 1440 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 1500 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 1560 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 1620 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 1680 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 1740 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 1800 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 1860 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 1920 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 1980 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 2040 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 2100 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 2160 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 2220 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 2280 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 2340 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 2400 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 2460 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 2520 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 2580 cccgtatcgt agttatctac acgacgggga gtcaggccac tatggatgaa cgaaatagac 2640 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 2700 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 2760 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 2820 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 2880 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 2940 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 3000 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 3060 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 3120 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 3180 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 3240 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 3300 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 3360 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 3420 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 3480 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 3540 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 3600 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 3660 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 3720 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 3780 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 3840 accatgatta cgccaagctt gcatggcggc cgcataactt cgtatagcat acattatacg 3900 aagttatgca tgcttatgaa gcgacgatgg aaaaagaatt tcatagccgt ctcagcagcc 3960 aaccgcttta agaaaatctc atcctccggg gcacttgatt atggtacccc aactactgct 4020 tctgaaaatc tatattttca aggtgaacta aaaactgctg ctttggctca acatgcgatt 4080 aaagctgatg cgcaacaaaa taacttcaac aaagatcaac aaagcgcctt ctatgaaatc 4140 ttgaacatgc ctaacttaaa cgaagcgcaa cgtaacggct tcattcaaag tcttaaagac 4200 gacccaagcc aaagcactaa cgttttaggt gaagctaaaa aattaaacga atctcaagca 4260 ccgaaagctg ataacaattt caacaaagaa caacaaaatg ctttctatga aatcttgaat 4320 atgcctaact taaacgaaga acaacgcaat ggtttcatcc aaagcttaaa agatgaccca 4380 agccaaagtg ctaacctatt gtcagaagct aaaaagttaa atgaatctca agcaccgaaa 4440 gcggataaca aattcaacaa agaataaggc gcgccacttc taaataagcg aatttcttat 4500 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa 4560 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca 4620 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat 4680 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac 4740 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca 4800 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt 4860 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag 4920 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc 4980 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct 5040 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta 5100 aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg 5160 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg 5220 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg 5280 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca 5340 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa 5400 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg 5460 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc 5520 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg 5580 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc 5640 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta 5700 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc 5760 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga 5820 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt 5880 tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa 5940 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt 6000 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa 6060 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata 6120 ctaacgccgc catccagtcg acgaactagt ataacttcgt atatggtatt atatacgaag 6180 ttatgcggcc gctgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg 6240 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg 6300 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc 6360 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata ccatcgtccg 6420 gatcaatgaa gtagatgagt ttactacctg tatatacttt tttttttgtc aaagtcactt 6480 ttatggcatt tccatgttga aggattagcc acagcactaa attttcagcg tatgattgct 6540 tttaaatatt taattttcat cgttttttaa ttaatattca aaacgattta atgccttcta 6600 catctgaagt caagctcata gactcgggtt gggtagctaa aactacatct aacacgaagg 6660 ggtatagcct tcttaaagta tactcatgta taaacgtttt gggttcatat ttttgggcca 6720 cgtggtttat aaaaattctt aactacacca ctcggtatcc cgcacccgtc tacgtttcta 6780 cgatttcgag ctaatattga gtaaagtgat tagcaaaaaa aataaaagta ctcgttgtcg 6840 gagatcaaga atttttctat tatctcatct aaaccacttt ctaaaagcga aaaacaaaat 6900 cgtaatatgc agcttgaatg ggcttccata gtttgaaaga aaaaccctag cagtactggc 6960 aagggagaca ttccttttac ctggctaatc gccatcttcc agcaggcgca ccattgcccc 7020 tgtttcacta tccaggttac ggatatagtt catgacaata tttacattgg tccagccacc 7080 agcttgcatg atctccggta ttgaaactcc agcgcgggcc atatctcgcg cggctccgac 7140 acgggcactg tgtccagacc aggccaggta tctctgacca gagtcatcct tagcgccgta 7200 aatcaatcga tgagttgctt caaaaatccc ttccagggcg cgagttgata gctggctggt 7260 ggcagatggc gcggcaacac cattttttct gacccggcaa aacaggtagt tattcggatc 7320 atcagctaca ccagagacgg aaatccatcg ctcgaccagt ttagttaccc ccaggctaag 7380 tgccttctct acacctgcgg tgctaaccag cgttttcgtt ctgccaatat ggattaacat 7440 tctcccaccg tcagtacgtg agatatcttt aaccctgatc ctggcaattt cggctatacg 7500 taacagggtg ttataagcaa tccccagaaa tgccagatta cgtatatcct ggcagcgatc 7560 gctattttcc atgagtgaac gaacctggtc gaaatcagtg cgttcgaacg ctagagcctg 7620 ttttgcacgt tcaccggcat caacgttttc ttttcggatc cgccgcataa ccagtgaaac 7680 agcattgctg tcacttggtc gtggcagccc ggaccgacga tgaagcatgt ttagctggcc 7740 caaatgttgc tggatagttt ttactgccag accgcgcgcc tgaagatata gaagataatc 7800 gcgaacatct tcaggttctg cgggaaacca tttccggtta ttcaacttgc accatgccgc 7860 ccacgaccgg caaacggaca gaagcatttt ccaggtatgc tcagaaaacg cctggcgatc 7920 cctgaacatg tccatcaggt tcttgcgaac ctcatcactc gttgcatcga ccggtaactt 7980 ttagattaac atttgtttaa atatgaaatt aggcaatgta acatactgca ggcaaatttt 8040 ggtgtacggt aagtaaattg gacatatgat ttaacaaagc gactataagt cagaaagtga 8100 gaatgagatt gaaataatta attcaactta ttcaattgat gatatgccag gattcctctt 8160 cctttatata tcttcaatgt ttcgattaac aggtccatta tccggttttt agtttcgctg 8220 cacattgccg aatgacaatc ggattccttt aagctttgct tttgccctgt tttctctccc 8280 cagtaatcgt tgggtgactg aaccatccag tagccgagag ttacgttttt cctgatttaa 8340 taaatatcca tatctatttg tctcgtggtg ttttacagca actaaccgaa attactattt 8400 tgcatcatcc aaccataaag agacaacaat aattctagaa aacatataac aagtgttttt 8460 ttctatttac gtctcatacg caaagtccac tttaccatgt tttggaaact tgtacccaac 8520 cattcatttt tcgattttgt aatgaggctc ttttcttcta acaatcattg aggtcattat 8580 catgtgtcct cacctattgt ttaaattgag gcttgcgcta ttcccaaaat tgtgggtatt 8640 attttcatat atcagtatct gcaatttcgg ttcaactccg gaagctcggt acccaactgt 8700 gggaatactc aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt 8760 ttctctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact 8820 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat 8880 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg 8940 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat 9000 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca 9060 tttcagcaat atatatatat atatatatat atatttcaag gatataccat tgtaatgtct 9120 gcccctaaga agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa 9180 gccattaagg ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa 9240 aatcatttaa ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg 9300 ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg 9360 ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg 9420 tacgccaact taagaccatg taactttgca tccgactctc ttttagactt atctccaatc 9480 aagccacaat ttgctaaagg tactgacttc gttgttgtta gagaattagt gggaggtatt 9540 tactttggta agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac 9600 accgttccag aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag 9660 ccaccattgc ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg 9720 agaaaaactg tggaggaaac catcaagaac gaatttccta cattgaaagt tcaacatcaa 9780 ttgattgatt ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata 9840 atcaccagca acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggctcc 9900 ttgggtttgt tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt 9960 ttgtacgaac catgccatgg ttccgctcca gatttgccaa agaataaggt caaccctatc 10020 gccactatct tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt 10080 aaagccattg aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta 10140 ggtggttcca acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc 10200 cttgcttaaa aagattctct ttttttgtga tatttgtaca taaactttat aaatgaaatt 10260 cataatagaa acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat 10320 atacgcaatc tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg 10380 taagcaaatt gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta 10440 atattgacaa ggaggagggc atcaca 10466 (pAW8_TAP"Times New Roman LbMluI(392),XbaI(798),ScaI(1014) Arial(e\SS Lb+ScaI(1111),NdeI(1192),XbaI(1214),ClaI(1429) Arial(|tDVt LbScaI ArialP(Wajj LbHPvuII(3677),HindIII(3856),EagI(3868),NotI(3868),KpnI(4008),HindIII(4363) Arial($+22q# Lb7AscI(4470),BglII(4706),PstI(4920),MluI(4953),NcoI(5093) Arial8(&&e Lb#NruI(5177),ClaI(5211),HindIII(5638) Arial[(Ajmj LbYScaI(5909),SalI(6138),SpeI(6146),NotI(6187),EagI(6187),EcoRI(6195),PvuII(6287),PacI(6572) Arial+ *(e Lb,ScaI(6830),ScaI(6955),ClaI(7208),EcoRV(7465) Arial+(e LbFSexAI(7584),BamHI(7657),NruI(7802),PstI(8031),NdeI(8065),HindIII(8252) Arialz(:::B^ LbXbaI(8435),KpnI(8693) Arial (Z Lb!ClaI(9267),KpnI(9366),EcoRV(9865) Arial$(|tlHl LbNcoI Arial&(8V5L2B6LBLbC("8 pGEX_3_primer"Times New RomanLbCɆ( AmpR_promoter"Times New RomanLbC(  Ampicillin"Times New RomanLbC(y ORF 3"Times New RomanLb(   pBR322_ori"Times New RomanLb( lac_promoter"Times New RomanLbC(M13_pUC_rev_primer"Times New RomanLbCɆ(M13_reverse_primer"Times New RomanLbCɆ(sORF 1"Times New RomanLbC("CloxP"Times New RomanLb(pHybLex_rev_primer"Times New RomanLb(kanR"Times New RomanLbC(8HM13_forward20_primer"Times New RomanLb(AWM13_pUC_fwd_primer"Times New RomanLbCɆ(IORF 1"Times New RomanLbC(#'ORF 2"Times New Roman Lbtest"Times New Roman (E6` Lbtest"Times New Roman( !r5 Lbtest"Times New Roman(